Classifications

• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
  • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
  • C07K14/575—Hormones
  • C07K14/66—Thymopoietins
  • C07K14/662—Thymopoietins at least 1 amino acid in D-form
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P37/00—Drugs for immunological or allergic disorders
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K38/00—Medicinal preparations containing peptides
• • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
  • Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
  • Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
  • Y10S930/00—Peptide or protein sequence
  • Y10S930/01—Peptide or protein sequence
  • Y10S930/18—Thymus derived hormone or factor; related peptides

Definitions

• thymus-dependent lymphocytes T cells
• the hydrophobic region can comprise 1 to 2 amino acids or their amides or esters, where S -Ile or S -Val as central area give the peptides a particularly good quality of action.
• the Alkanoy can in particular in A l rest to be a formyl Hexanoylrest, the aroyl group is benzoyl which, methoxy or chlorine is optionally substituted by methyl, the Aralkanoylrest phenacetyl, Cinnamoyl, dihydrocinnamoyl, phenoxyacetyl or phenylthiacetyl, where the aryl radical may be unsubstituted or substituted by methyl, methoxy or chlorine.
• Alkyl or aralkyloxycarbonyl is preferably ethyl, isobutyl, tert-butyl, benzyl, 4-methylbenzyl, 4-methoxybenzyl or 4-chlorobenzyloxycarbonyl.
• the amino acid representing Y with a hydrophobic side chain can in particular be Ala, Val, Leu, Ile, Met, Phe, Pro, Tyr, Phg (C-phenylglycine), and also a hydrophobically substituted amino acid such as Ser (Bu), T hr (Bu t ), C y s ( Bu t ), Cys (Et), Cys (Bzl), Glu (OBu t ), Asp (OBu t ), Glu (NH-Bu t ), Glu (NH-Et), Lys (Boc ), Orn (Boc), Tyr (Bu t ), Tyr (Me), Phe (Cl), Tyr (Cl), furthermore alkyl.
• a hydrophobically substituted amino acid such as Ser (Bu), T hr (Bu t ), C y s ( Bu t ), Cys (Et), Cys (Bzl), Glu (OBu t ), Asp (OBu t ), Glu
• alkyl amides are n-alkyl or branched alkyl amides such as isopropyl, isobutyl, tert-butyl, 3-methylbutyl or 3-ethylbutylamide; as aralkylamides, for example benzyl- or phenethylamide, which can be substituted in the core and / or in the side chain by 1 to 2 methyl groups.
• aralkylamides for example benzyl- or phenethylamide, which can be substituted in the core and / or in the side chain by 1 to 2 methyl groups.
• the same alkyl, aryl or aralkyl radicals can also be present in an ester bond.
• the peptides according to the invention are similar to. a pentapeptide Arg-Lys-Asp-Val-Tyr recently described in Science 204 (1979), page 1309, a partial sequence of thymopoietin, which is addressed as the region of this peptide which is said to be responsible for the biological action. Interestingly, this sequence corresponds to the above-mentioned sequence basic-basic-acid-hydrophcb-hydrophobic, which thus appears as a superordinate principle for peptides of the thymic activity mentioned.
• the compounds according to the invention also include tri- and tetrapeptide derivatives, the effect of which is in many cases the same or even increased compared to a pentapeptide of the sequence Lys-Lys-Glu-Val-Val produced according to the invention.
• the invention further relates to a process for the preparation of the peptides mentioned, which is characterized in that amino acid sequences of the formula A - B - S - X - Y in which A, B, S, X and Y have the meaning given in claim 1, is based on methods of peptide synthesis.
• the peptides according to the invention have a significantly longer lifespan than natural thymus peptides.
• S is an acidic D-amino acid such as D-glutamic acid or D- ⁇ -aminoadipic acid are particularly noteworthy.
• the compounds according to the invention can be used for the treatment of immunodeficiencies, viral and fungoid, and also chronic bacterial infections, autoimmune diseases, and for the therapy of diseases which are caused by cells with immunologically relevant changes in the cell membrane characteristics (e.g. tumor cells).
• the invention also relates to the use of the peptides mentioned in general to influence the maturation of T lymphocytes.
• the residue is triturated in succession with water, 1N citric acid, saturated sodium bicarbonate solution and water and dried.
• To remove the protective groups dissolve in 25 ml of trifluoroacetic acid and precipitate after 40 min. the crude pentapeptide derivative with ether.
• the trifluoroacetate obtained is converted into the acetate by stirring with a strongly basic ion exchanger in the acetate form in 50% methanol, which is present as a resinous semi-solid mass after filtering off the exchanger and distilling off the solvent.
• Example 1 The procedure of Example 1 is followed, except that 5.6 g of H-Val-Tyr-NH 2 , prepared according to Chem. Ber. 97 (1964), page 1197, and after removal of the protective groups and purification, receives 9.0 g of the title compound as acetate.
• H-Val-OIpr ⁇ HCl prepared in a known manner by esterifying L-valine with HCl / isopropanol, are dissolved in 20 ml of dimethylacetamide with the addition of 4.0 ml of N-ethylmorpholine.
• symmetrical Boc-Val anhydride is prepared from 13.1 g of Boc-Val-OH and 6.6 g of DCC in 50 ml of dimethylacetamide at 0 °.
• the two solutions are combined, allowed to come to room temperature, stirred for a further 4 hours at about 22 ° and the solvent is distilled off in vacuo. The residue is taken up in 150 ml of ethyl acetate and extracted three times with 20 ml of water.
• the Boc compound is dissolved in 60 ml of trifluoroacetic acid. After 40 min. precipitate with ether / petroleum ether (1: 1), which is digested thoroughly with the same mixture. After drying in vacuo over KOH, the yield is 6.1 g, mp. 172-174 ° C. Elemental analysis (N, F ) correct.
• Example 5 B The Boc residue is split off analogously to Example 5 B). Out booty 3.1 g. Almost uniform in the DC.
• Isoleucine and HCl / isopropanol are prepared in a known manner from isoleucine isopropyl ester hydrochloride. 2.1 g of this compound are reacted analogously to Example 5 C) with 14.4 g of Adoc-Arg (Adoc) 2 -Arg (Adoc) 2 -Glu (OBU t ) -OH ⁇ 2 H 2 0 and worked up. Expanded 6.2 g. Amino acid analysis: Glu 1.00, Ile 0.95, Arg 2.06.
• the tert-butyl group is split off with trifluoroacetic acid analogously to Example 1, converted into the acetate and the compound is purified by chromatography on Sephadex (R) LH 20. Yield 2.9 g, amino acid analysis: Glu 1.00, Val 1.89, Arg 2.01.
• Example 12 The procedure of Example 12 is followed, but ZD-Lys (Boc) -ONSu is used. The crude product is catalytically hydrogenated in 90% acetic acid on Pd. Then, the procedure for cleavage of t Boc and Bu, for conversion into the acetate and subsequent purification in analogy to Examples 12 and 1. Yield 6.0 g. Amino acid analysis: Glu 1.00, Val 1.89, Lys 0.99, Arg 1.02.
• the resinous residue is dissolved in 100 ml of methanol and hydrogenated catalytically on Pd with titration with 2N HCl in methanol at pH 4. After filtering off the catalyst, the mixture is brought to dryness, the solid residue is taken up in 50 ml of dimethylformamide and 4.0 g of ethyl pentachlorophenyl carbonate (Bull. Soc. Chim. France 23 (1900), page 818) are added. The mixture is stirred overnight, the solvent is distilled off in vacuo and the residue is digested with ether. The procedure for removing the protective groups, converting them to acetate and for cleaning is analogous to Example 1. Yield 4.1 g. Amino acid analysis: Glu 1.00, Val 1.92, Lys 2.02.
• Example 15 The procedure of Example 15 is followed, except that 1.1 g of succinic anhydride are used instead of the phthalic anhydride and the procedure is as described in Example 15. Yield 6.1 g. No free ⁇ -amino group can be recognized by the dansyl method.
• the active ester is prepared from 2.31 g of Boc- ⁇ -aminocaproic acid, 1.15 g of HONSu and 2.02 g of DCC in 10 ml of dimethylformamide, and the precipitated dicyclohexylurea is filtered off, and the filtrate is combined with the solution of 5.2 g of Arg-Glu (OBu t ) -Val -Val-OMe, 2HCl, (Example 11 B) in 25 ml of dimethylformamide and adds 1.03 ml of N-ethylmorpholine.
• the change in concentration in the eluate due to the emerging peptides is determined via the refractive index (differential refractometer). Yield after lyophilizing the fraction, redissolving in 5% acetic acid and lyophilizing again 0.6 g.
• Amino acid analysis Glu 1.00, Val 1.88, ⁇ -Guanidocaproyl not determined, ⁇ -Aminocaproyl ⁇ 0.02, Arg 1.01.
• the compound obtained according to B) is hydrogenated catalytically on Pd in 80 percent methanol.
• the catalyst is filtered off and the solution is brought to dryness in vacuo. Trituration of the residue with ether leads to the chromatographically pure compound.
• the ethyl acetate phase is then extracted with 150 ml of saturated NaHCO 3 solution, K 2 SO 4 / KHSO 4 solution, saturated NaHCO 3 solution and water, dried over Na 2 SO 4 and concentrated.
• Example 21 B 31 g of H-Val-Val-CBu t ⁇ HCl are condensed analogously to Example 21 B) with 37 g of Z-Glu (OBzl) -OH in dimethylformamide in the presence of 13.5 g of HOBt and 12.8 ml of N-ethylmorpholine using 22 g of DCC.
• catalytic hydrogenation is carried out as described in Example 21 C).
• the ether-insoluble compound is isolated and is uniform by thin layer chromatography. Yield 51 g.
• Pd / carbon catalyst is added to a solution of 65 g (0.134 mol) of Z-Val-Tyr (Bu t ) -OMe in 300 ml of methanol and the mixture is passed with stirring and with the addition of about 2N methanolic hydrochloric acid at pH 4.5 (autotitrator ) Hydrogen through the solution until methanolic hydrochloric acid is no longer absorbed.
• the catalyst is then filtered off and the filtrate is concentrated. The residue is triturated with ether, the substance dissolving and crystallizing after standing overnight at 4 °. The crystals are filtered off and dried over P 2 0 5 .
• the still wet precipitate is dissolved in the required amount of ethyl acetate and the adhering water is separated off in the separating funnel.
• the ethyl acetate phase is dried over Na 2 SO 4 and evaporated almost to dryness.
• the peptide is precipitated from the concentrated solution with petroleum ether.
• the mixture is cooled to 4 ° C. and the precipitate is filtered off and dried over P 2 O 5 .
• the substance (44 g) is boiled with 1000 ml of ethyl acetate, mixed with 1000 ml of petroleum ether and cooled. The precipitate is filtered off and dried. Educ. 43.15 g.
• the substance is boiled with 450 ml of methanol. The mixture is allowed to cool to room temperature and suction filtered, washed with methanol and dried in vacuo.
• the substance is chromatographed on silica gel in the solvent mixture methylene chloride: methanol: water: acetic acid such as 90: 15: 2: 2. Educ. 2.78 g (63%).
• the amorphous residue was processed without characterization.
• the precipitate is filtered off and washed with water.
• the still water-moist product is distributed between 150 ml of ethyl acetate and 150 ml of water.
• the still wet precipitate is dissolved in 200 ml of ethyl acetate.
• the adhering water is separated.
• the ethyl acetate phase is shaken out once more with 50 ml of water, dried over Na 2 SO 4 and concentrated. It is overturned from ethyl acetate / petroleum ether.
• Example 30 F Analogously to Example 30 F are Z-Glu (OBu t) -Ile-T yr (Bu t) -O-cyclohexyl catalytically hydrogenated in 200 ml of methanol, 5 g (6.65 mmol). Educ. 4.3 g (99%).
• Example 30 H Analogously to Example 30 H are hydrogenated catalytically 4.0 g (4:08 mmol) of Z-Lys (Boc) -Glu (OBu t) -Ile-Tyr (Bu t) -O-cyclohexyl.
• Example 30 L Analogously to Example 30 L, 3 g (about 2.2 mmol) of Z-Arg (Z 2 ) -Lys-Glu-Tle-Tyr-O-cyclohexyl in 150 ml of 90 percent. Acetic acid catalytically hydrogenated and worked up. Educ. 1,675 g (76%). For further purification, 800 mg of a hydroxypropylated cross-linked dextran gel in 90 percent. Chromatographed methanol.
• Example 30 K Analogously to Example 30 K are treated 3.1 g (ca. 2.2 mmol) of Z-Arg (Z 2) -Lys (Boc) -Glu (OBu t) -Val-Tyr (Bu t) -On-hexyl with 90 percent trifluoroacetic acid. Educ. 2.42 g (93%).
• Example 30 Analogously to Example 30 are treated K 5.74 g (4.2 mmol) of Z-Arg (Z 2) -Lys (Boc) -Glu (OBu t) -Val-Tyr (Bu t) -On-butyl with 90 percent trifluoroacetic acid. Educ. 4.99 g.
• Example 30 K Analogously to Example 30 K are 6.75 g (5 mmol) of Z-Arg (Z 2) -Lys (Boc) -Glu (OBu t) -Val-Tyr (Bu t) -O-CH (CH 3) 2 90-percent with Trifluoroacetic acid treated. Educ. 5.66 g.
• the catalyst is filtered off, the filtrate is concentrated and the residue is dissolved in water.
• the solution is chromatographed on a weakly basic ion exchanger in acetate form (30 x 2 cm). Elution with water.
• the peptide-containing eluate is freeze-dried and, for further purification on a hydroxypropylated cross-linked dextran gel in 90 percent. Chromatographed methanol. Educ. 430 mg amino acid analysis: Asp (1.00), Val (1.1), Tyr (0.9) Lys (1.09), Arg (0.98), content of peptide base: approx. 78%.
• Example 38 A Analogously to Example 38 A are 4:04 g (12 mmol) ZD-Glu (OBu t) -OH condensed with 4.64 g (12 mmol) H-Val-Tyr (Bu t) -OMe ⁇ HCl in 25 ml of dimethylformamide. Educ. 6.72 g.
• Example 38 B Analogously to Example 38 B, 5.4 g (8 mmol) of ZD-Glu (OBu t ) -Val- T yr (Bu t ) -OMe are catalytically hydrogenated. The residue is dried in a high vacuum. Educ. 4.57 g (100%). The substance is amorphous.

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• 1979
  • 1979-09-22 DE DE19792938420 patent/DE2938420A1/de not_active Withdrawn
• 1980
  • 1980-08-29 AT AT80105158T patent/ATE6857T1/de not_active IP Right Cessation
  • 1980-08-29 EP EP80105158A patent/EP0025897B1/fr not_active Expired
  • 1980-08-29 DE DE8080105158T patent/DE3067271D1/de not_active Expired
  • 1980-09-16 ES ES495088A patent/ES495088A0/es active Granted
  • 1980-09-19 ZA ZA00805830A patent/ZA805830B/xx unknown
  • 1980-09-19 AU AU62550/80A patent/AU541852B2/en not_active Ceased
  • 1980-09-19 CA CA000360613A patent/CA1182448A/fr not_active Expired
  • 1980-09-21 IL IL61098A patent/IL61098A/xx unknown
  • 1980-09-22 JP JP13276480A patent/JPS5653640A/ja active Pending
• 1981
  • 1981-11-09 US US06/319,267 patent/US4420424A/en not_active Expired - Fee Related

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